Abstract: Objective To investigate the histocompatibility between a new type of endotamponade hydrogel substitute and human retinal Müller cells in vitro. Design Experiment study. Participants Human retinal Müller cell line (MIO-M1). Methods Human retinal Müller cells were passaged and cultured until stable growth, and then divided into three groups, which were respectively treated with a certain concentration of balanced salt solution (BSS), new type of Hydrogel extract, and perfluorocarbon liquid (PFCL) for cultivation. The morphology of cells in each group was observed after 24 h. The cells were cultured for 1 h, 4 h, 12 h, 24 h, 48 h, and 72 h, respectively. The cell viability was measured using MTT assay at different time points. Flow cytometry was used to detect the cell apoptosis, and the concentration of IL (interleukin)-6 in the supernatant was measured using ELISA at different time points. Meanwhile, expression of intracellular reactive oxygen species (ROS) was identified by immunofluorescence after 24 h of culture. Main Outcomes Measures Cell morphology, cell viability, IL-6 concentration, cell apoptosis rate, ROS concentration. Results After co-culturing the new hydrogel extract and Müller cells for 72 h, compared with the group of BSS, there was no decrease in cell viability or cell survival rate, no significant change in cell morphology, no significant increase in the secretion level of IL-6 and intracellular ROS after 24 h. In addition, compared with the group of BSS, after co-culturing PFCL and Müller cells for 12 h, the secretion level of IL-6 was increased (P=0.0105), the cell viability started to decrease after 24 h (P=0.0147), and the cells in contact with PFCL shrank, the number of the cells decreased, and the arrangement of cells was loose. Meanwhile, the analysis of apoptosis at different time points showed that the late apoptosis and necrosis rate of Müller cells increased after 24 h (P=0.0201). The level of intracellular ROS significantly increased after 24 h（P<0.0001）. Conclusions The new type of endotamponade hydrogel substitute has good histocompatibility with human retinal Müller cells in 72 h. Exposing Müller cells to PFCL for 12 h can increase the secretion level of IL-6. After 24 hours, the cell viability decreased and the level of apoptosis increased, which may be related to the oxidative stress mechanism activated by ROS in cells. (Ophthalmol CHN, 2023, 32: 509-515)

Key words: Müller cells, endotamponade substitute, perfluorocarbon liquid, hydrogel, artificial vitreous body